Biomedical subjects

M J Delgado

Publications and source records attributed to M J Delgado.

At least 55 records · Page 3Linked to original sources

PubMed1995

Central administration of beta-endorphin increases food intake in goldfish: pretreatment with the opioid antagonist naloxone.

The effect of intraperitoneal or intracerebroventricular beta-endorphin administration on food intake has been studied in satiated goldfish. Food intake was evaluated at different time intervals after injections, 0-2, 2-8 and 0-8 h. The 0.1 and 1 micrograms doses of beta-endorphin intracerebroventricularly administered induced an increase in food intake during the first 2 h postinjection, while no modifications on feeding were observed in the next 6 h. These same doses of beta-endorphin used increased cumulative food intake at 8 h postinjection. In contrast, intraperitoneal injection of 1 micrograms of beta-endorphin did not modify food intake in any of the studied time intervals. Naloxone, an opioid antagonist, attenuated the beta-endorphin-induced feeding increase. These results suggest that opioids may play a role in modulation of feeding central regulation, acting via opioid receptors in goldfish.

Analysis of Variance↗

PubMed1995

Seasonal changes in thyroid activity in male and female frog, Rana perezi.

Plasma T3 and T4 levels, and thyroid free (f) and bound (b) thyroid hormones contents, were determined by radioimmunoassay in adult male and female Rana perezi over 1 year. Plasma T3 levels show a significant seasonal cycle in both male and female frogs, increasing from January to peak in July (71-74 pg/ml) to decrease in August-October. Plasma T4 concentrations also varied seasonally, and sex differences were apparent. Basal T4 levels from the end of summer to the end of winter and higher values (682-651 pg/ml) in spring and early summer occur in female frogs; males have lowest levels in October (59.78 +/- 13.92 pg/ml) and higher levels during the rest of the year, except for a significant decrease in April. The thyroid content of fT3 and fT4 displayed similar seasonal patterns, with peak values in April and July-August. Sex differences are again present. The thyroid contents of bT3 are high between March and July in both sexes, with values in females being greater than those in males. The bT4 content follows a similar seasonal pattern in males and females and shows a minimum in March and a maximum in summer. Thyroid releasing capacity seems to change depending on the season and sex.

Animals↗

PubMed1995

CRF effect on thyroid function is not mediated by feeding behavior in goldfish.

In the present study we examined the effects of acute corticotropin-releasing factor (CRF) administration and refeeding treatment on glucose levels and thyroid hormones (plasma levels and thyroid contents) in 48-h food-deprived goldfish. Central CRF administration (2 micrograms) decreased food intake and the thyroid T3 free fraction, without significantly modifying either thyroid hormones bound fractions (T3 and T4) or plasma glucose levels. Subsequently, we tested whether CRF affects thyroid activity by itself, or whether this effect is mediated by CRF-induced feeding reduction. CRF treatment in fasted fish reduced thyroid-free T3 and increased thyroid-free T4, which could be mediated by a decreased intrathyroidal 5'-monodeiodinase activity. These data suggest a CRF effect upon thyroid activity independent of feeding reduction. On the other hand, refeeding after 48-h fasting caused a significant increase in thyroid free T4 content and plasma thyroid hormone levels. Thus, a relationship between nutritional status and thyroid function, which could overlap with CRF effects, cannot be discarded. Plasma glucose levels were only significantly modified by refeeding, which seems to be the signal triggering the increase in glucose titers. Our results support the existence of both CRF-thyroid activity and nutritional status-thyroid function interactions in goldfish.

Animals↗

PubMed1995

Differential characteristics and regulation of arylamine and arylalkylamine N-acetyltransferases in the frog retina (Rana perezi).

Arylamine N-acetyltransferase activity (A-NAT: E.C.2.3.1.5) from Rana perezi retina was studied using p-phenetidine as specific substrate. Enzyme characteristics and regulation were compared with respect to the arylalkylamine N-acetyltransferase (AA-NAT: E.C.2.3.1.87) from the same tissue. A-NAT activity is distributed in both neural retina and choroid-pigmented epithelium complex, showing a 10-fold higher specific activity in neural retina. In contrast, AA-NAT activity is restricted to neural retina. Subcellular localization in neural retina indicated that both enzymatic activities are in the supernatant fraction (39,000 g, 20 min). p-Phenetidine acetylation was linear as a function of the neural retina amount in the assay (1/16 to 1 retina), and it is insensitive to phosphate buffer pH in the range 6.5-8.4. A-NAT kinetic showed a hyperbolic shape for both cosubstrates. Kinetic constants were KM = 11.2 microM, Vmax = 0.49 nmol/h/mg prot. for p-phenetidine (50 microM acetyl-CoA), and KM = 113.4 microM, Vmax = 3.1 nmol/h/mg prot. for acetyl-CoA (5 mM p-phenetidine). The additivity test for both enzymatic activities in retina homogenates demonstrated that both acceptor amines do not compete for the catalytic sites. Serotonin addition in the assay modifies differentially the kinetic characteristics of both enzymes. Serotonin acted as a strong mixed inhibitor, mainly competitive in nature (competitive Ki = 18.1 microM; non-competitive Ki = 1.9 mM) for AA-NAT. However, it acted as a weak inhibitor with respect to A-NAT, mainly non-competitive, (competitive Ki = 5.7 mM; non-competitive Ki = 8.7 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Changes in the fatty acid profile of plasma and adipose tissue in rats after long-term ethanol feeding.

The effect of chronic ethanol feeding on the fatty acid composition of plasma and abdominal adipose tissue in rats was studied. Animals were maintained on a 30% ethanol solution in drinking water for 3 and 5 months. Control rats were given water. Caloric intake was similar in control and ethanol-fed rats at the end of the experimental period. However, a decrease in body weight was observed in rats that had consumed ethanol. Palmitoleic (16:1n7) and oleic (18:1n9) acids increased markedly, and linoleic acid (18:2n6) decreased in the plasma and in the adipose tissue of ethanol-fed rats with respect to control rats. After 3 months of ethanol ingestion, long-chain polyunsaturated fatty acids were reduced both in plasma and adipose tissue. When ethanol was administered for 5 months, only plasma long-chain polyunsaturated fatty acids of the n-3 series were decreased. This suggest that changes induced by ethanol ingestion in essential fatty acid metabolism is less pronounced when ethanol feeding is maintained for a long period of time.

Adipose Tissue↗

PubMed1995

Characterization of the cycHJKL genes involved in cytochrome c biogenesis and symbiotic nitrogen fixation in Rhizobium leguminosarum.

Mutants of Rhizobium leguminosarum bv. viciae unable to respire via the cytochrome aa3 pathway were identified by the inability to oxidize N,N'-dimethyl-p-phenylenediamine. Two mutants which were complemented by cosmid pIJ1942 from an R. leguminosarum clone bank were identified. Although pea nodules induced by these mutants contained many bacteroids, no symbiotic nitrogen fixation was detected. Heme staining of cellular proteins revealed that all cytochrome c-type heme proteins were absent. These mutants lacked spectroscopically detectable cytochrome c, but cytochromes aa3 and d were present, the latter at a higher-than-normal level. DNA sequence analysis of complementing plasmids revealed four apparently cotranscribed open reading frames (cycH, cycJ, cycK, and cycL). CycH, CycJ, CycK, and CycL are homologous to Bradyrhizobium japonicum and Rhizobium meliloti proteins thought to be involved in the attachment of heme to cytochrome c apoproteins; CycK and CycL are also homologous to the Rhodobacter capsulatus ccl1 and ccl2 gene products and the Escherichia coli nrfE and nrfF gene products involved in the assembly of c-type cytochromes. The absence of cytochrome c heme proteins in these R. leguminosarum mutants is consistent with the view that the cycHJKL operon could be involved in the attachment of heme to apocytochrome c.

Amino Acid Sequence↗

PubMed1995

Alterations in food intake and thyroid tissue content by corticotropin-releasing factor in Tinca tinca.

The present experiments test the effects of intracerebroventricular injections of ovine corticotropin-releasing factor (1 microgram) on food intake, plasma glucose levels and thyroid function at 8 h postinjection in tench. Food intake and thyroid triiodothyronine (T3) content were significantly decreased after CRF treatment. Thyroid thyroxine (T4) content and plasma glucose levels were not modified by this neuropeptide. The present results suggest that CRF plays a role on food intake regulation and thyroid gland activity in tench.

Animals↗

PubMed1994

Ontogeny of ocular serotonin N-acetyltransferase activity daily rhythm in four anuran species.

The profile of daily ocular serotonin N-acetyltransferase (NAT) rhythm has been examined during the development of four anuran amphibians in different environments and of varying phylogenetic origins: Xenopus laevis, Discoglossus pictus, Rana perezi, and Bufo calamita. A significant day/night rhythm of ocular NAT with the acrophase at midnight was first detected at stages 43/44 and 25 in X. laevis and R. perezi, respectively. Total enzymatic activity increased with body weight throughout development, although both mean levels and amplitudes of NAT activity per milligram of protein significantly decreased; this attentuation of rhythm was more significant in postmetamorphic froglets, and the acrophase was delayed about 1 hr. There were no significant differences in daily NAT profiles before and after metamorphosis in D. pictus and B. calamita. The amplitude of the NAT rhythm was maximal (> 10-fold nocturnal values) in Xenopus compared with other species (2- to 3-fold).

Animals↗

PubMed1994

Serotonin N-acetyltransferase activity as a target for temperature in the regulation of melatonin production by frog retina.

The adaptive mechanisms of serotonin N-acetyltransferase (NAT) activity in the regulation of melatonin synthesis in frog retina in the face of chronic and acute temperature changes have been investigated. We performed thermal acclimation experiments to test different environmental temperatures at two seasons of the year (summer and winter), followed by the set-up of an eyecup culture system to investigate the acute effects of temperature on NAT activity and melatonin production daily rhythms. Low temperature induced a significant increase in NAT activity, independent of both the time of the photocycle (midday or midnight) and the season of the year (winter or summer). Acute cold-induced stimulation of NAT activity may be associated with lower decreases in the enzyme synthesis rate, rather than decreases in the degradation rate. In contrast, acclimation to warm temperature (25 degrees C) stimulated ocular melatonin production. Nocturnal melatonin production in eyecups cultured at 25 degrees C was significantly higher than in eyecups cultured at 5 degrees C. We suggest that this discrepancy in thermal regulation of melatonin synthesis can be justified by a seasonal variation in serotonin content within the photoreceptor cells, which determines the thermal response of melatonin production through changes in NAT kinetics.

Acclimatization↗

PubMed1994

Comparative effects of intestinal absorption of folic acid and methyltetrahydrofolic acid in chronic ethanol-fed rats.

This study concerns in vivo folic acid and methyltetrahydrofolic acid (MTHF) absorption by the whole intestinal surface after 20 weeks of 30% ethanol ingestion in drinking water. The results were compared with control rats fed ad libitum. The total intestinal serosal areas were similar in ethanol-fed and control rats. Significant increases in intestinal length, and decreases in tissue wet and dry weights were found in ethanol-fed rats. Serum folic acid concentrations were significantly less in the animals which had ingested ethanol than in the control rats. Intestinal folic acid absorption was significantly increased at lower substrate concentrations (0.5 and 1 microM), while no difference was observed at 2.5 microM in the ethanol-fed rats. Folic acid absorption relative to tissue wet weight showed significant increases at all tested concentrations in the ethanol-fed rats. Intestinal MTHF absorption showed no significant changes at 0.5 microM MTHF concentration, and an increase was observed in the absorption values at 1 and 2.5 microM concentrations in the ethanol-fed rats. When expressed as tissue wet weight, MTHF absorption values in ethanol-fed rats increased at 1 and 2.5 microM but did not differ at 0.5 microM substrate concentrations. The above results indicate compensatory responses in the folic acid and MTHF intestinal absorption after chronic ethanol ingestion. These effects are observed when the whole intestinal surface is evaluated.

Animals↗

PubMed1993

The inhibition by indoleamines (tryptamine and serotonin) of ocular serotonin-N-acetyltransferase from Rana perezi is temperature-dependent.

Temperature effects on ocular serotonin N-acetyltransferase (NAT) kinetics characteristics from Rana perezi have been studied with respect to tryptamine and serotonin as substrates. Monoamine oxidase (MAO) activity does not interfere in NAT assay at acceptoramine concentrations used in NAT kinetics characterization from R. perezi retina. NAT shows an inhibition by high substrate (serotonin) concentration, which is temperature-dependent. NAT follows the Michaelis-Menten equation at low temperature, whereas at high temperatures (> 10 degrees C) an inhibition by serotonin is observed. This inhibition of NAT activity by serotonin could act as an amplification mechanism to increase daily melatonin rhythm amplitude in the retina of ectotherms.

Animals↗

PubMed1993

Serotonin N-acetyltransferase (NAT) activity and melatonin levels in the frog retina are not correlated during the seasonal cycle.

The seasonal pattern of daily serotonin N-acetyltransferase (NAT) and melatonin rhythms in the lateral eye of Rana perezi has been characterized to assess any seasonal relationship between enzymatic activity and melatonin production and to evaluate the photothermal responses of such rhythms. In winter there was no daily rhythm either in NAT activity or in melatonin production, although the highest values of NAT activity and lowest melatonin concentrations were measured at this time. Ocular melatonin showed a high-amplitude diurnal rhythm during the summer, whereas a low-amplitude rhythm was observed in spring and autumn. Diurnal melatonin levels were temperature-independent; however, nocturnal melatonin production significantly increased with temperature. Ocular NAT was always higher at lower than at higher temperatures (two- to threefold), both at night and during the day. As suggested for pineal melatonin production in other poikilotherm species, photoperiod controls the duration of the melatonin nocturnal rise, whereas temperature regulates the amplitude of the daily melatonin rhythm in the frog retina. The lack of correlation between daily NAT and melatonin rhythms in the frog retina may result from the high thermal sensitivity of the ocular NAT and a temperature-dependent inhibition substrate of NAT activity.

Animals↗

PubMed1993

Role of corticotropin-releasing factor (CRF) as a food intake regulator in goldfish.

The effect of intraperitoneal or intracerebroventricular corticotropin-releasing factor (CRF) administration on food intake has been studied in goldfish after 24 h of food deprivation. Food intake was evaluated at different time periods after injection, 0-2, 2-8, and 0-8 h. The 1 and 2 micrograms doses of CRF intracerebroventricularly administered induced a reduction in food intake during the first 2 h, although an increased feeding was observed in the next 6 h. The higher dose of CRF used in this study (3.3 micrograms) increased cumulative food intake at 8 h postinjection. However, intraperitoneal injection of 1 microgram CRF did not modify food intake in any of the studied intervals. These results suggest that CRF may play a role in central regulation of feeding behavior intake in goldfish, and show that CRF effects are time- and dose-dependent.

Animals↗

PubMed1992

Thermal sensitivity and effect of temperature acclimation on ocular serotonin N-acetyltransferase activity in Rana perezi.

The thermal sensitivity and the response to thermal acclimation of the serotonin N-acetyltransferase (NAT) activity in frog ocular tissue were studied. The ocular NAT shows a positive thermal modulation for both cosubstrates (tryptamine and acetyl-CoA). The higher NAT activity in the cold-acclimated group (4 degrees C) with respect to the warm-acclimated one (24 degrees C) implies a partial thermal compensation of the enzyme. The present results suggest that frog ocular NAT response to temperature entails a modulation of thermal sensitivity of the enzyme rather than changes in enzyme concentration.

Acclimatization↗

PubMed1992

Corticotropin-releasing factor stimulates metamorphosis and increases thyroid hormone concentration in prometamorphic Rana perezi larvae.

Attempts to identify a hypothalamic molecule that stimulates thyrotropin (TSH) secretion from amphibian pituitary have been unsuccessful to date. The effects of mammalian (ovine and human) corticotropin-releasing factor (CRF) on the thyroid function of prometamorphic (Taylor & Kollros stages XI-XVII) (Taylor and Kollros, 1946) Rana perezi larvae were studied. Chronic treatments with both ovine and human CRF (oCRF, hCRF) stimulated metamorphosis while delaying larval growth. Chronic hCRF (1 microgram) administration induced 3.2- and 5.3-fold increases in whole body concentration of thyroxine (T4) and triiodothyronine (T3), respectively. In contrast, the 0.5-microgram dose of hCRF stimulated a significant (3.4-fold) increase in whole body concentration of T4 but not of T3. Histological studies of the thyroid gland revealed a 22% increase in the number of follicles per section as a result of the chronic treatment with oCRF (1 microgram). Acute oCRF (2 micrograms) treatment induced a significant increase in T4 concentration at 4 hr (1.3-fold) and 8 hr (2.3-fold) postinjection. T3 concentration was not altered. These results support previous reports and lead us to conclude that a CRF-like peptide, and not TRH, is involved in the regulation of thyroid activity in anuran amphibians during metamorphosis.

Animals↗

PubMed1992

Characterization of serotonin N-acetyltransferase in the lateral eye of the green frog Rana perezi: protective action of EGTA.

The kinetics of serotonin N-acetyltransferase (NAT) from the lateral eye of Rana perezi have been characterized. NAT from ocular tissue reached maximal activity at a phosphate buffer concentration of 250 mM and a pH of 6.5. Reaction linearity was highly conserved within the homogenate fraction range tested (0.033-0.33). The time course of ocular NAT reaction showed a high linearity at 25 and 35 degrees C. Km and Vmax estimations for acetyl-CoA at a 10 mM tryptamine concentration were 63.3 microM and 4.42 nmol/h per eye, respectively. Regardless of the acceptor amine (tryptamine or serotonin), the Km was not affected by the acetyl-CoA concentration (50 or 250 microM), whereas the Vmax was significantly increased at a 250 microM acetyl-CoA concentration. Ocular NAT showed a higher affinity for serotonin (Km = 20.7 microM) than for tryptamine (Km = 48-60 microM); Vmax, however, was similar for both substrates. Acetyl-CoA does not protect ocular NAT; in contrast, the use of EGTA (greater than or equal to 4 mM) in the assay is essential to protect the enzyme because NAT in ocular crude homogenate shows rapid inactivation. This result suggests that intracellular calcium levels are involved in the NAT inactivation mechanisms in frog ocular tissue.

Acetyl Coenzyme A↗

PubMed1992

Role of environmental temperature and photoperiod in regulation of seasonal testicular activity in the frog, Rana perezi.

To analyze the role of environmental temperature and photoperiod in the regulation of the annual testicular cycle in Rana perezi, we performed experiments combining high (25 +/- 1 degrees C) or low (6 +/- 1 degrees C) temperature and different photoperiod regimens (18L:6D, 12L:12D, and 6L:18D (hours light:hours dark)) during three phases of the reproductive cycle: winter stage (December) and prebreeding (February) and postbreeding (May, June) periods. Low temperature and short photoperiod in winter induced the arrest of the maturation phase of spermatogenesis and the activation of primary spermatogonia proliferation and spermiohistogenesis. Rana perezi testis responded to long days stimulus in winter, even at low temperature, with induction of the maturation phase of the cycle. Exposure of male frogs to either high temperature or long photoperiod induced a decrease in testosterone levels in winter. During the prebreeding period, an increase in environmental temperature caused a reduction in testosterone, and a lengthening in photoperiod produced the opposite effect. Photoperiod had no effect on testosterone levels during the postbreeding period, but low temperature increased testosterone plasma levels. These results suggest that both temperature and photoperiod effects can vary seasonally, depending on the phase of the annual reproductive cycle in R. perezi.

Animals↗

PubMed1990

mRNA transcription determines the lag period for the induction of pineal melatonin synthesis in the Syrian hamster pineal gland.

The nocturnal pattern of Syrian hamster pineal melatonin synthesis is characterized by a 6-8 h lag period, followed by a late-night, short-duration peak in both N-acetyltransferase (NAT) activity and melatonin content. Administration of cycloheximide (20 mg/kg body weight) given either at the time of lights out or 4 h into the dark phase to Syrian hamsters blocked the nocturnal increase in both pineal NAT activity and melatonin content. Actinomycin D (5 mg/kg body weight) prevented the nocturnal increase in both constituents only when it was administered at darkness onset, being significantly less effective when injected after 4 h of dark exposure. Reinduction of melatonin production by isoproterenol (2 mg/kg body weight) administration to acutely light-exposed animals during late darkness was prevented by cycloheximide, but not by actinomycin D administration. The results suggest that whereas Syrian hamster pineal melatonin production requires protein synthesis both early and late in the dark phase, the transcription of a putative NAT-related mRNA, which occurs only during the early night, seems to determine the lag period in melatonin synthesis and pineal responsiveness to beta-adrenergic receptor agonist stimulation.

Acetyltransferases↗